Characterization of MADS-domain transcription factor complexes in Arabidopsis flower development

Abstract

Floral organs are specified by the combinatorial action of MADSdomain transcription factors, yet the mechanisms by which MADSdomain proteins activate or repress the expression of their target genes and the nature of their cofactors are still largely unknown. Here,we showusing affinity purification andmass spectrometry that five major floral homeotic MADS-domain proteins (AP1, AP3, PI, AG, and SEP3) interact in floral tissues as proposed in the "floral quartet"model. In vitro studies confirmed a flexible composition of MADSdomain protein complexes depending on relative protein concentrations and DNA sequence. In situ bimolecular fluorescent complementation assays demonstrate that MADS-domain proteins interact duringmeristematic stages of flower development. By applying a targeted proteomics approach we were able to establish a MADS-domain protein interactome that strongly supports a mechanistic link between MADS-domain proteins and chromatin remodeling factors. Furthermore, members of other transcription factor families were identified as interaction partners of floral MADS-domain proteins suggesting various specific combinatorial modes of action.